LOCUS       sequence-85-expo        4638 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4638)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1995
REFERENCE   2  (bases 1 to 4638)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pPD93_51
FEATURES             Location/Qualifiers
     source          1..4638
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             386..406
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(449..477)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1225..1245
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(2212..2234)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2334..2353
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2431..2449)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2517..2621
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2622..2690,2691..3482)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2840..2859)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3653..4241
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4142..4161
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4395..4412
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4529..4550
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4565..4570,4571..4588,4589..4595)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4603..4619
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     4608..4630
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(4627..4638,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(4627..4638,1..5)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 atgaccatga ttacgccaag cttgcatgcc tgcaggtcga ctctagagga tccccaattc
       61 ttgagacttc cactatcttc tctggcaccc tccactacgc gtaggcgtct ctcgcttcgt
      121 gtattcccgg gaagccgggt tcccgtctct cccgccgctg ccgctgccgc acacagcttt
      181 acacctcgta gaatccccaa agaggggcgt ggcttgcggg tgccaacatc ctcctgccga
      241 ggaagaagca ggcactcatc actcgcatca tcaacctcgg gattggccaa aggacccaaa
      301 ggtatgtttc gaatgatact aacataacat agaacatttt caggaggacc cttggagggt
      361 accgagctca gaaaaaatga ctgctccaaa gaagaagcgt aaggtaccgg tagaaaaaat
      421 gagtaaagga gaagaacttt tcactggagt tgtcccaatt cttgttgaat tagatggtga
      481 tgttaatggg cacaaatttt ctgtcagtgg agagggtgaa ggtgatgcaa catacggaaa
      541 acttaccctt aaatttattt gcactactgg aaaactacct gttccatggg taagtttaaa
      601 catatatata ctaactaacc ctgattattt aaattttcag ccaacacttg tcactacttt
      661 cacttatggt gttcaatgct tctcgagata cccagatcat atgaaacggc atgacttttt
      721 caagagtgcc atgcccgaag gttatgtaca ggaaagaact atatttttca aagatgacgg
      781 gaactacaag acacgtaagt ttaaacagtt cggtactaac taaccataca tatttaaatt
      841 ttcaggtgct gaagtcaagt ttgaaggtga tacccttgtt aatagaatcg agttaaaagg
      901 tattgatttt aaagaagatg gaaacattct tggacacaaa ttggaataca actataactc
      961 acacaatgta tacatcatgg cagacaaaca aaagaatgga atcaaagttg taagtttaaa
     1021 catgatttta ctaactaact aatctgattt aaattttcag aacttcaaaa ttagacacaa
     1081 cattgaagat ggaagcgttc aactagcaga ccattatcaa caaaatactc caattggcga
     1141 tggccctgtc cttttaccag acaaccatta cctgtccaca caatctgccc tttcgaaaga
     1201 tcccaacgaa aagagagacc acatggtcct tcttgagttt gtaacagctg ctgggattac
     1261 acatggcatg gatgaactat acaaatagca ttcgtagaat tccaactgag cgccggtcgc
     1321 taccattacc aacttgtctg gtgtcaaaaa taataggggc cgctgtcatc agagtaagtt
     1381 taaactgagt tctactaact aacgagtaat atttaaattt tcagcatctc gcgcccgtgc
     1441 ctctgacttc taagtccaat tactcttcaa catccctaca tgctctttct ccctgtgctc
     1501 ccacccccta tttttgttat tatcaaaaaa cttctcttaa tttctttgtt ttttagcttc
     1561 ttttaagtca cctctaacaa tgaaattgtg tagattcaaa aatagaatta attcgtaata
     1621 aaaagtcgaa aaaaattgtg ctccctcccc ccattaataa taattctatc ccaaaatcta
     1681 cacaatgttc tgtgtacact tcttatgttt tttacttctg ataaattttt ttgaaacatc
     1741 atagaaaaaa ccgcacacaa aataccttat catatgttac gtttcagttt atgaccgcaa
     1801 tttttatttc ttcgcacgtc tgggcctctc atgacgtcaa atcatgctca tcgtgaaaaa
     1861 gttttggagt atttttggaa tttttcaatc aagtgaaagt ttatgaaatt aattttcctg
     1921 cttttgcttt ttggggtttc ccctattgtt tgtcaagatt tcgaggacgg cgtttttctt
     1981 gctaaaatca caagtattga tgagcacgat gcaagaaaga tcggaagaag gtttgggttt
     2041 gaggctcagt ggaaggtgag tagaagttga taatttgaaa gtggagtagt gtctatgggg
     2101 tttttgcctt aaatgacaga atacattccc aatataccaa acataactgt ttcctactag
     2161 tcggccgtac gggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     2221 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     2281 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc
     2341 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag
     2401 gagaaaatac cgcatcaggc ggccttaagg gcctcgtgat acgcctattt ttataggtta
     2461 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     2521 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     2581 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     2641 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     2701 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     2761 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     2821 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     2881 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     2941 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     3001 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     3061 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     3121 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     3181 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     3241 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     3301 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     3361 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     3421 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     3481 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     3541 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     3601 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     3661 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     3721 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     3781 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
     3841 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     3901 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     3961 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     4021 aactgagata cctacagcgt gagcattgag aaagcgccac gcttcccgaa gggagaaagg
     4081 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     4141 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     4201 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     4261 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     4321 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     4381 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac
     4441 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
     4501 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc
     4561 aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat
     4621 ttcacacagg aaacagct
//
